Soybeans and soybeans products having high palmitic acid content

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Iowa State University Patents

Iowa State University Research Foundation, Inc.

2-11-1997

Soybeans and soybeans products having high

palmitic acid content

Walter R. Fehr

Iowa State University, wfehr@iastate.edu

Earl G. Hammond

Iowa State University

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http://lib.dr.iastate.edu/patents

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Sciences Commons

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Plant Pathology Commons

This Patent is brought to you for free and open access by the Iowa State University Research Foundation, Inc. at Iowa State University Digital Repository. It has been accepted for inclusion in Iowa State University Patents by an authorized administrator of Iowa State University Digital Repository. For more information, please contactdigirep@iastate.edu.

Recommended Citation

Fehr, Walter R. and Hammond, Earl G., "Soybeans and soybeans products having high palmitic acid content" (1997).Iowa State University Patents. 139.

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Soybeans and soybeans products having high palmitic acid content

Abstract

A method is described for producing soybean varieties and lines exhibiting palmitic acid contents of at least

about 18.0% up to 30.0% or more. The novel soybean lines are obtained from a soybean seed designated

A1937NMU-85 and its descendants, particular desirable progeny resulting from the cross of A1937NMU-85

with ElginEMS-421, and further with the cross of selected progeny with A89-259098.

Keywords

Agronomy

Disciplines

Agricultural Science | Agriculture | Agronomy and Crop Sciences | Life Sciences | Plant Pathology | Plant

Sciences

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United States Patent [191

Fehr et al.

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5,602,311

Feb. 11, 1997

[11] Patent Number:

[45] Date of Patent:

[54] SOYBEANS AND SOYBEANS PRODUCTS

HAVING HIGH PALMITIC ACID CONTENT

[75] Inventors: Walter R. Fehr; Earl G. Hammond,

both of Ames, Iowa

[73] Assignee: Iowa State University Research

Foundation, Inc., Ames, Iowa

[21] Appl. No.: 376,466

[22] Filed: Jan. 20, 1995

Related US. Application Data

[63] Continuation of Ser. No. 180,114, Jan. 12, 1994, abandoned,

which is a continuation of Ser. No. 839,328, Feb. 20, 1992, abandoned, which is a continuation-in-part of Ser. No.

461,341, Jan. 5, 1990, abandoned.

[51] Int. Cl.6 ... .. A01H 5/00; AOlH 5/10;

C12N 15/01

[52] US. Cl. ... .. 800/200; 800/230; 800/250;

800/DIG. 26; 800/DIG. 69; 435/1721;

426/601

[58] Field of Search ... .. 800/200, 230,

800/250, DIG. 26, DIG. 69; 426/601; 435/172.1

[56] References Cited

PUBLICATIONS

Kuhr et a1 1987 Release Notice for N85-2124, N 85-2131 and N85-2176.

Carter et a1 1988 Release Notice for C1726 and C1727.

Mounts et al 1987 In Soybean Improvement, Production and

Uses; Agronomy Monograph No. 16; Wilcox (ed) pp.

819-845 and 860—865.

Neumunz 1978 J. Am. Oil Chem. Soc. 55: 396A-398A.

Weihrauch et a1 1977 Food Technology 31:80-85 and 91.

Graef et a1 1985 J. Am. Oil Chem Soc. 64:773-775. Wilcox et a1 1984 J Am Oil Chem Soc. 61297-100.

Wilson et al 1981 Crop Sci 21:788-791.

Burton et al 1983 Crop Sci 23:744-747.

Erickson et a1 1988 Crop Sci 28:644-646.

Bubeck et a1 1989 (May-Jun.) Crop Sci 29:652-656. Wilcox 1989 (5 Mar.) In Conf. Mundial de Investigation en Soja; pp. 28-39.

Primary Examiner—Patricia R. Moody

Attorney, Agent, or Firm—Burns, Doane, Swecker & Mathis, L.L.P.

[57] ABSTRACT

A method is described for producing soybean varieties and lines exhibiting palmitic acid contents of at least about

18.0% up to 30.0% or more. The novel soybean lines are

obtained from a soybean seed designated A1937NMU-85

and its descendants, particular desirable progeny resulting

from the cross of A1937NMU-85 with ElginEMS-42l, and further with the cross of selected progeny with A89-259098.

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5,602,311

1

SOYBEANS AND SOYBEANS PRODUCTS HAVING HIGH PALMITIC ACID CONTENT

This is a continuation of application Ser. No. 08/180,114,

?led on 12 Jan. 1994 (now abandoned), which is a continu

ation of application Ser. No. 07/839,328, ?led on 20 Feb.

1992 (now abandoned), which is a continuation-impart of

Ser. No. 07/461,341, ?led 5 Jan. 1990 (now abandoned).

FIELD OF THE INVENTION

This invention relates to novel soybean seeds and prod ucts of soybean seeds, such as soybean oil, and, more

particularly, to soybeans seeds and products characterized

by extremely high levels of palmitic acid.

BACKGROUND OF THE INVENTION

Soybean seeds represent perhaps the most signi?cant

oilseed in the world. Soybean oil makes up approximately 28% of the world supply of fats and oils, has been consid ered to be the major vegetable oil produced and consumed in the United States, and more than 90% of the soybean oil

is used in food products (World Soybean Research Confer

ence IH Proceeding, Shibles, R. (Ed.) 1985).

Soybeans thus represent a signi?cant world-wide food

source, providing an excellent source of protein. As such,

soybeans represent potential alternatives to meats.

Tofu and soymilk are two principal food products derived from soybean seeds. More than one billion people in China and SoutheastAsia, it has been stated, rely on tofu as a major food protein source. (Proc. Int. Soya Protein Food Conf., American Soybean Assoc., p. 35 (1970)). Soyrnilk is simi larly an important source for food protein.

One application for which soybean oil may be used is the

production of plastic fats (e.g.—shortenings and marga

rines). Such plastic fats are made with a matrix of solid fats whose interstices are ?lled with liquid oil. Solid fats can

crystallize in several forms with different melting points and physical properties. The forms are commonly designated alpha, beta‘ and beta, with the beta form having the highest melting point and the greatest stability. Forms other than

these three may also be present. The beta‘ form generally has the properties that are most usually desired in a plastic fat. If the solid portion of the plastic fat contains about 15%

or more of palmitic acid and the rest is stearic acid, it will

stabilize in the beta‘ form. If the ratio of stearic/palmitic is

higher, then the fat may convert to the beta form with its less

desirable physical structure.

Soybean 'oil will most usually contain a level of about 10% palmitic acid or so. Accordingly, if such soybean oil is hydrogenated and made into a plastic fat, it will likely

crystallize in the beta form. To prevent this, an oil such as

cottonseed or palm oil that is richer in palmitic acid is blended with the soybean oil.

The use of either palm oil or cottonseed oil presents some di?iculties. Some users thus consider palm oil to be unde

sirable based upon perceived health considerations. On the

other hand, cottonseed oil is generally available only in

limited amounts at a higher price than that of soybean oil. It would accordingly be highly desirable to be able to provide

soybean varieties having su?iciently elevated palmitic acid

contents so that plastic fats can be produced such that such

products will stabilize in the beta‘ form.

10 15 25 30 35 45 50 55 60 65

2

Further, some producers for some plastic fat applications believe that soybean oil having a palmitic acid content in the

range of about 13% or 14% to about 16% or so is preferred.

For such applications, it would be highly desirable to be able

to provide soybean varieties having a sut?ciently high level

of palmitic acid to use for blending with soybean varieties having more conventional palmitic acid contents to provide the desired intermediate range of palmitic acid content.

The palmitic acid levels in soybean seed oil range from

9.3% to 17.4% within the world collection (Erickson et al.,

Journal of Heredity, 79, p. 465, 1988). The Erickson et al. article reports the inheritance of altered palmitic acid per centages in two soybean mutants, C1726 and C1727. The level of palmitic acid in C1727 reported averages 17.3% palmitic acid in comparison to 11.5% in the oil of the parent

cultivar “Century.”

Despite the clear need for soybeans having a level of palmitic acid above that present in the world collection at the present time, this objective still remains to be achieved.

SUMMARY OF THE INVENTION

It has been discovered that a mutant line obtained from the

parent variety Asgrow A1937 provides a population of soybean seeds exhibiting high levels of palmitic acid. The

palmitic acid concentration capable of being obtained is

about 18% of the total fatty acid present, even up to 20% or

more.

Further, it has been discovered that crossing the Asgrow

A1937 mutant with a second mutant provides a population

of soybean seeds exhibiting signi?cantly higher levels of palmitic acid than have been capable of being achieved. More particularly, soybean seeds having palmitic acid con

tents above about 20%, and, more preferably, of at least about 25% or more, have been achieved. Still further, it has

been discovered that soybean seeds having palrnitio acid

contents of 30.0% or more may be obtained by crossing

selected progeny with line A89-259098.

If desired, the soybeans of the present invention can‘be

used as a donor parent in a backcrossing program with any

desired commercial cultivar as a recurrent parent to isolate

a commercial variety having desirable seed yield and other agronomic characteristics in addition to the high level of

palmitic acid.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENT

According to one aspect of the present invention, the

novel soybean seeds and plants characterized by palmitic

acid contents of at least 18% were obtained by preparation

of a mutant line developed by treating the cultivar Asgrow A1937 with nitroso methyl urea (NMU). The mutation procedure utilized will be described in detail hereinafter in the Examples. The mutant line has been designated A1937NMU-85.

Pursuant to a further aspect of the present invention,

crossing of the parent mutant line A1937NMU-85 with a second mutant line, ElginEMS-421, has been found to

provide a population of soybean seeds with palmitic acid contents signi?cantly above that of the parent line Asgrow A1937. The preparation of the mutant line, ElginEMS-421, will be described in conjunction with the Examples.

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5,602,311

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technique. Standard hybridization techniques are, of course,

well known and may be utilized. As an illustrative example,

hybridization techniques are disclosed in Fehr, Principles of

Cultivar Development, Vol. 1, Theory and Technique, Chap

ter 13, pp. 156—l64, Macmillan Publishing Company, New York, 1987, which hybridization techniques are herein incor~

porated by reference.

Progeny from the crossing of Al937NMU-85 and Elg

inEMS-42l yielded soybean seeds wherein the palmitic acid content is greater than 18%, preferably greater than about 20%, and more preferably, greater than about 25%.

The fatty acid composition was determined by gas-liquid

chromatography using the method as generally outlined in

Graef et al. (Crop Sci., 25:l076—l079, 1985). Thus, in general, the method comprises (1) crushing the seed sample,

(2) putting the crushed sample into a test tube with a hexane

solvent and extracting the oil into the hexane, (3) the fatty

acids in the oil are converted to their methyl esters using sodium methoxide and methanol, (4) water is added to

inactivate the sodium methoxide catalyst, and (5) the methyl

esters, which ?oat to the top of the water layer, are diluted

with hexane and become the sample that is introduced into the column of the gas chromatography apparatus.

As may be appreciated, this general methodology may be employed and speci?c aspects changed to lessen the time needed as desired. For example, the stationary phase

selected for the columns will dictate the temperature at which the sample can be introduced.

None of the speci?cs utilized, e.g.——capillary versus

packed columns, are considered to affect to any appreciable extent the results obtained for an analysis. Rather, such

speci?cs affect the time required for sample preparation and

analysis.

The percentages of the fatty acids set forth herein, unless

otherwise designated, thus are on a weight basis and refer to the percentage of the methyl ester of palmitic acid or other

fatty acid compared to the total methyl esters of the fatty acid composition in the sample being analyzed. This can also be taken as the weight percentage of the fatty acid itself because the difference between the palmitic acid content and

that of its methyl ester as determined in the gas chromatog raphy technique described herein is so minimal as may be

ignored, as is commonly done in this ?eld.

The gas chromatography techniques described herein are

routinely used for analysis of the fatty acid composition of

soybeans. The experimental error is considered to be within

the range of from about 1 to 5% or so, depending upon the

magnitude of the peak. For example, with a relatively large

peak indicative of an oleic acid content of 50% or so, the experimental error may be as low as about 1% of the value, viz.——50i0.5%. At the other extreme, a small peak indica tive of a stearic acid content of 4.0% may have an experi mental error of about 5.0% of the value, viz.—4.0% i0.2. A

palmitic acid content of about 20% lies in the middle, with the expected error being about 2—3% of the reported value,

viz.—-20%i0.4 or 0.6%.

As may be appreciated, the palmitic acid levels of the

soybeans of the present invention set forth herein were

obtained from soybeans grown in Iowa and Puerto Rico.

Growth under climatic conditions cooler or warmer may

result in a somewhat altered fatty acid composition. How ever, while the speci?c results may vary somewhat depend

ing upon the speci?c growing conditions experienced, the

progeny of the present invention will be characterized by extremely high palmitic acid contents relative to other soybean lines grown under similar conditions.

20 25 45 50 55 65

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Progeny exhibiting the desired high palmitic acid trait can

be crossed with other progeny to provide a population of

soybean seeds having extremely high palmitic acid contents.

It can be expected that crosses utilizing the more desirable

progeny should be capable of providing lines having palm

itic acid contents up to about 30.0% or so.

Indeed, soybean lines having palmitic acid contents of

30.0% or more have been obtained by crossing selected

progeny with line A89-259098. Parent line A89-259098 is

described in our copending application, Ser. N o. 07/643,277,

the disclosure of said line being herein incorporated by

reference. It can thus be expected that crosses utilizing the more desirable progeny should be capable of providing lines

having palmitic acid contents up to about 35.0% or so.

Further, progeny can be crossed, if desired, with other

progeny, or with any other soybean line or cultivar to yield a soybean cultivar having the desired seed yield or other desired agronomic traits as well as the desired high palmitic

acid trait. Self-pollination of selected progeny may likewise yield lines having characteristics desired for some applica

tions.

Any hybridization technique may be used, and many are

known as has been described herein. For example, the

selection of progeny having the desired high palmitic acid trait can be obtained by conducting backcrossing with a commercial variety until a desirable commercial variety has

been isolated. Backcrossing techniques are known, as dis

closed in Fehr, Principles of Cultivar Development, Vol. 1, Theory and Technique, Chapter 28, pp. 360—376, the dis—

closure of which is herein incorporated by reference. As one example, backcrossing using the desired F2 seeds

obtained by natural self-pollination of the F1 plants could be

carried out as follows:

(1) Plant F1 seeds obtained by crossing a parent with the

desired high palmitic acid trait to the desired commer

cial cultivar (recurrent parent). Sample F2 seeds from F1 plants are analyzed for fatty acid concentration, and

seeds with the desired high palmitic acid content are

planted for backcrossing.

(2) Cross-pollinate the desired commercial cultivar (recurrent parent) with an F2 plant having the high

palmitic acid content.

(3) Plant the BCIF1 seeds and obtain BC1F2 seeds by natural self-pollination. Sample BCIF2 seeds are ana

lyzed for fatty acid concentration, and those displaying

the high palmitic acid trait are backcrossed to the

recurrent parent.

(4) The backcross and selection procedure herein described (Step 3) can be repeated until lines with the

desired high palmitic acid composition and agronomic

performance are recovered. It is believed that four of these backcross cycles should serve to transfer the high

palmitic acid trait to the desired cultivar (viz.—recur rent parent), although the number of such cycles can be

fewer, or more, as is desired. The result is the produc tion of a soybean line quite similar to the commercial

cultivar except having the desired high palmitic acid

content.

Any commercial cultivar (recurrent parent) desired may

be employed for backcrossing. Factors such as, for example, seed yield, geographical area, and many others, as is known, will generally dictate the cultivar selected from the several hundred commercial cultivars available.

The following Examples are illustrative, but not in limi tation, of the present invention. The gas chromatography

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5

two decimal points (i.e.—“0.00”). As reported herein, the

fatty acid values are set forth to one decimal point. Values of 6 or more in the second decimal point were raised (e.g.-— 4.29 is reported herein as 4.3), values of 4 or less are ignored

(e.g. —4.24 is reported as 4.2), values of 5 are raised if the ?rst decimal is odd (cg-4.15 is reported as 4.2) and ignored if even e.g.—4.25 is reported as 4.2).

EXAMPLE 1

This Example describes the preparation of the mutant line A1937NMU-85.

Mutant line A1937NMU-85 was obtained from nitroso

methyl urea (NMU) treatment of the parent variety Asgrow A1937. In May, 1985, 2,500 seeds of A1937 were soaked in

2.5 L distilled water in a 6 L ?ask for 9 hours at room temperature. The ?ask was aerated for the 9 hours of soaking. The water was drained from the ?ask, and 2.5 L of

2.5 rnM NMU in 0.1 molar [“M”] phosphate buffer at pH 5.5

were added. The seeds were soaked with aeration for 3 hours, the solution was drained and the seeds were rinsed twice with distilled water. Treated seeds were placed in

containers to prevent drying and transported to the Agricul tural Engineering and Agronomy Research Center near Ames, Iowa. The seeds were planted 2.5 cm deep in moist

soil within 4 hours after the last rinse. The soil was watered

regularly to keep it moist until seeding emergence. The

properties of the mutant seed and their progeny were evalu

ated beginning with the M4 generation.

A similar number of seeds was harvested from each of the

M1 (?rst mutant generation) plants in the population to obtain 2,000 M2 seeds. A random sample of 1,000 of the

second generation M2 seeds from the population was planted in October at the Iowa State University-University of Puerto Rico soybean nursery at Isabela, Puerto Rico. About 2,000 M3 seeds were obtained by harvesting a similar number of seeds from each M2 plant. In February, 1,000 M3 seeds were planted in Puerto Rico. About 2,000 M4 seeds were obtained

by harvesting a similar number of seeds from each M3 plant.

In May, 1,000 M4 seeds were planted at Ames. Five hundred

M4 plants were harvested individually from the population,

and a 10-seed sample from selected plants was analyzed by gasaliquid chromatography to determine the fatty acid com position. M5 progeny of selected plants were planted in Puerto Rico in February, 1987; and the results con?rmed the

unique fatty acid composition of the M4 parent plant.

A 10-seed sample of the M4 plant from which A1937NMU-85 originated and its parent was analyzed by

gas-liquid chromatography, and the results are set forth in

Table I:

TABLE I

Fatty Acid Composition

Palmitic Stearic Oleic Linoleic Linolenic Seed Acid Acid Acid Acid Acid

Identi?cation (16:0) (18:0) (18:1) (18:2) (18:3)

A1937NMU—85 19.8 3.6 17.4 51.3 7.7 A1937 Parent 12.3 3.8 18.7 57.2 8.0

M5 Progeny from the M4 plant A1937NMU-85 and the A1937 parent were analyzed by gas-liquid chromatography,

and the results are set forth in Table II:

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TABLE II

Fatty Acid Composition

Palrnitic Stearic Oleic Linoleic Linolenic Seed Acid Acid Acid Acid Acid

Identi?cation (16:0) (18:0) (18:1) (18:2) (18:3)

Al937NMU-85 20.1 3.8 18.5 49.9 7.6

A1937 Parent 12.1 4.6 27.1 50.1 6.1

Seeds of the soybean parent A1937NMU-85 have been deposited under the terms of the Budapest Treaty at the American Type Culture Collection (ATCC) at 12301 Parklawn Drive, Rockville, Md. 20852, USA. More spe ci?cally 2,500 seeds of A1937NMU-85 were deposited on Jun. 18, 1996 and have been assigned ATCC Accession No. 97618.

EXAMPLE 2

This Example describes the preparation of the mutant line

ElginEMS-42l.

Mutant line ElginEMS-42l was obtained from ethyl methane sulfonate (EMS) treatment of the parent variety Elgin. In May, 1984, 2,500 seeds of Elgin were soaked in 2.5

L distilled water in a 6 L ?ask for 9 hours at room

temperature. The ?ask was aerated for the 9 hours of soaking. The water was drained from the ?ask, and 2.5 L of

0.025 molar EMS in 0.1M phosphate bu?’er at pH 7 were

added. The seeds were soaked for 9 hours, the solution was drained and the seeds were rinsed twice with distilled water.

Treated seeds were placed in containers to prevent drying

and transported to the Agricultural Engineering and

Agronomy Research Center near Ames, Iowa. The seeds

were planted 2.5 cm deep in moist soil within 4 hours after the last rinse. The soil was watered regularly to keep it moist

until seedling emergence. The properties of the mutant seed and their progeny were evaluated beginning with the M2

generation.

A similar number of seeds was harvested from each of the

M1 (?rst mutant generation) plants in a population to obtain

2,000 M2 seeds for each population. A random sample of

1,000 of the second generation M2 seeds from the population was planted in February at the Iowa State University

University of Puerto Rico Soybean nursery at Isabela,

Puerto Rico. Five hundred M2 plants were harvested indi vidually from the population, and a l0~seed sample from

selected plants was analyzed by gas-liquid chromatography

to determine the fatty acid composition. M3 progeny of the mutant plant were planted in Puerto Rico in November, 1986, and the results con?rmed the unique fatty acid com

position of the M2 parent plant.

Table HI sets forth the analysis of the 10-seed sample of

the M2 plant from which ElginEMS-42l originated as well

as that of its parent:

TABLE III

Fatty Acid Composition

Palrnitic Stearic Oleic Linoleic Linolenic Seed Acid Acid Acid Acid Acid

Identi?cation (16:0) (18:0) (18:1) (18:2) (18:3)

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Table IV sets forth the analysis of M3 progeny from the

M2 plant E1ginEMS-421 and that of its parent:

TABLE IV

Fatty Acid Composition

Palmitic Stearic Oleic Linoleic Linolenic Seed Acid Acid Acid Acid Acid

Identi?cation (16:0) (18:0) (18:1) (18:2) (18:3)

ElginEMS-42l 16.9 4.1 16.8 50.2 11.9 Elgin Parent 11.6 4.2 17.8 55.5 10.9

Seeds of the soybean parent Elgin EMS-421 have been deposited under the terms of the Budapest Treaty at the American Type Culture Collection (ATCC) at 12301 Parklawn Drive, Rockville, Md. 20852, U.S.A. More spe ci?cally 2,500 seeds of Elgin EMS-421 were deposited on Jun. 18, 1996 and have been assigned ATCC Accession No.

97617.

EXAMPLE 3

This Example describes the crossing of Al937NMU-85 and ElginEMS-421 to obtain the soybean lines of the present

invention characterized by high palrnitic acid contents.

Crosses were made between individual plants of

A1937NMU-85 and ElginEMS~42l at the Agricultural Engineering and Agronomy Research Center near Ames, Iowa, in the summer of 1987. The hybrid F1 seeds obtained from the ditferent plant-to-plant crosses were kept separate, and were designated AX4659 through AX4663 and

AX4676.

The F1 seed was planted in the Iowa State University University of Puerto Rico nursery at Isabela, Puerto Rico, in October, 1987. F2 seeds were obtained by natural self

pollination. Each F1 plant was harvested individually, and

the F2 seeds of each were maintained as a separate subpopu lation.

Forty F2 seeds from each of the six crosses were planted in Puerto Rico in February, 1988. F3 seeds were obtained by natural self-pollination. F2 plants were harvested individu

ally. A 5-seed sample from each F2 plant was analyzed for

fatty acid composition by gas chromatography.

Table V summarizes the analysis of the fatty acid com

position of the F3 seeds from individual F2 plants:

TABLE V

Fatty Acid Composition

Palmitic Stearic Oleic Linoleic Linolenic

Seed Acid Acid Acid Acid Acid

Identi?cation (16:0) (18:0) (18:1) (18:2) (18:3)

AX4659-2-3 21.3 4.1 15.9 50.4 8.2 AX4659-3-7 25.9 3.7 14.2 47.9 8.3 AX4659-3-8 23.9 3.7 17.5 46.6 8.2 AX4660-3-4 26.1 4.5 16.2 44.9 8.3

AX4663-5-2 25 .9 4.4 17.7 44.4 7.5

AX4663-5-4 26.8 4.1 12.5 48.2 8.3 AX4676—2-1l 23.6 3.6 15.7 48.7 8.4 A1937NMU—85 19.0 3.8 15.7 53.2 8.2 ElginEMS-421 17.4 4.0 14.6 53.0 10.9

Twelve F3 seeds from each selected F2 plant were planted in the Iowa State University-University of Puerto Rico nursery in October, 1988. F4 seeds were obtained by natural

self-pollination. Each F3 plant was harvested individually. A 30 35 40 45 50 60 65

8

lO-seed sample from each F3 plant was analyzed for fatty

acid composition by gas chromatography.

Table VI summarizes the analysis of the fatty acid com

position of the F4 seeds from individual F3 plants:

TABLE VI

Fatty Acid Composition

Palmitic Stearic Oleic Linoleic Linolenic Seed Acid Acid Acid Acid Acid

Identi?cation (16:0) (18:0) (18:1) (18:2) (18:3)

AX4659-2-3-9 27.8 3.8 13.9 44.1 10.3 AX4659»3-7-7 27.4 3.5 15.1 44.4 9.6 AX4659»3-8-6 27.4 3.4 16.1 42.8 10.3

AX4660-3-4-4 27.7 3.8 15.6 43.2 9.7 AX4663-5-2-9 28.0 3.9 13.1 43.0 12.0

AX4663-5-4—5 28.3 4.0 12.4 43.8 11.5

AX4676-2-11-8 28.2 3.7 12.1 44.3 11.7 A1937NMU-85 19.6 3.9 17.5 50.1 8.9 ElginEMS-421 16.6 4.0 19.8 49.0 10.6

Thirty F4 seeds from each selected F3 plant were planted at the Agricultural Engineering and Agronomy Research

Center near Ames, Iowa, in May, 1989. F5 seeds were

obtained by natural self-pollination. Each F4 plant was

harvested individually. A S-seed sample from each of 10 F4 plants tracing to an F3 plant were analyzed for fatty acid

composition by gas chromatography.

Table VII summarizes the analysis of the fatty acid

composition of the F5 seeds from individual F4 plants, the

parents, and C1727:

TABLE VII

Fatty Acid Composition

Palrnitic Stearic Oleic Linoleic Linolenic

Seed Acid Acid Acid Acid Acid

Identi?cation (16:0) (18:0) (18:1) (18:2) (18:3)

AX4659-2-3-9 25.7 3.8 14.6 46.2 9.6

AX4659-3-7-7 26.1 3.9 14.0 45.2 10.7

AX4659-3-8-6 25.0 4.5 20.4 42.1 7.9

AX4660-3-4-4 26.6 4.0 15.1 44.6 9.6 AX4663-5-2-9 26.4 4.1 15 .7 43.0 10.7 AX4663-5-4-5 25.6 4.2 16.6 44.0 9.6 AX4676-2-11-8 25.2 4.0 17.4 44.3 9.1

A1937NMU-85 18.8 5.0 20.7 48.3 7.2

ElginEMS-421 15.7 4.1 19.7 50.7 9.8

C1727 15.6 3.6 19.2 53.2 8.4

EXAMPLE 4

This Example describes the crossing of AX4663-5-4-5

with A89-259098, a high stearic acid line, to obtain a

soybean line according to the present invention character ized by an exceptionally high palrnitic acid content.

Parent line AX4663-5-4-5 was obtained as described in

Example 3. Parent line A89~259098 was obtained as

described in our copending application, Ser. No. 07/643,277. The hybrid F1 seeds obtained from the cross were designated

AX7016-AX7019 .

F1 seeds of the cross were planted in February, 1990, in the Iowa State University-University of Puerto Rico nursery at Isabela, Puerto Rico. F2 seeds were obtained by natural

self-pollination. Each F1 plant was harvested individually,

and the F2 seeds of each were maintained as separate

subpopulations.

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5,602,311

9

the embryonic axis (approximately one-third of the seed) was analyzed for fatty acid composition by gas chromatog

raphy.

Table VIII summarizes the analysis of the fatty acid

composition of an F2 seed having the desired high palmitic

acid content and that of the parent lines:

TABLE VIII

Fatty Acid Composition

Palmitic Stearic Oleic Linoleic Linolenic Seed Acid Acid Acid Acid Acid

Identi?cation (16:0) (18:0) (18:1) (18:2) (18:3)

AX7017-1-3 30.4 5.2 10.1 42.6 11.7

AX4663-5-4-5 28.6 4.0 11.9 43.0 12.4

A89-259098 8.0 30.3 21.5 34.6 5.6

What is claimed is:

1. A method for producing a soybean plant that yields mature seed having an elevated endogenous palmitic acid content comprising:

(a) crossing a ?rst soybean parent plant that is

Al937NMU-85 having ATCC Accession No. 97618, with a second soybean parent plant that is Elgin EMS

421 having ATFCC Accession No. 97617;

(b) obtaining hybrid soybean seeds from the cross and

germinating said seeds to produce segregating popula

tions of soybean plants that are allowed to self-polli nate; and

(c) selecting a plant of step (b) that yields mature seed following self pollination having an endogenous palm

itic acid content in the oil thereof in an amount higher

than the palmitic acid of the parent plant of step (a) having the higher endogenous palmitic acid content

when the parent plants and the o?spring of the cross are

grown under the same conditions, and wherein the

elevated palmitic acid content of the selected offspring

of the cross is a heritable trait.

2. A soybean plant which upon self pollination yields

mature seed that exhibits an elevated palmitic acid content

and is the product of crossing a ?rst soybean parent that is A1937NMU-85 having ATCC Accession No. 97618, with a

second soybean parent that is Elgin EMS-421 having ATCC

Accession No. 97617, and wherein the soybean plant result

ing from said cross yields mature seed that provides an

endogenous oil comprising palmitic acid in an amount higher than the endogenous palmitic acid content of the parent having the higher palmitic acid content when the

parents plants and the offspring of said cross are grown

under the same conditions, and wherein the elevated palm

itic acid content of the offspring of the cross is a heritable trait.

3. A mature soybean seed formed by the self pollination of the soybean plant according to claim 2 and the descen

15

25

35

45

50

10

dants thereof that retains said elevated palmitic acid content

when grown under the same conditions.

4. A method according to claim 1 wherein said soybean plant that is selected in step (c) contains an endogenous

palmitic acid content in the oil of the mature seed formed

thereon of at least about 18% by weight as determined by

gas chromatography.

5. A method according to claim 1 wherein said soybean plant that is selected in step (0) contains an endogenous

palmitic acid content in the oil of the mature seed formed

thereon of greater than about 20% by weight as determined

by gas chromatography.

6. A method according to claim 1 wherein said soybean plant that is selected in step (c) contains an endogenous

palmitic acid content in the oil of the mature seed formed

thereon of greater than about 25% by weight as determined

by gas chromatography.

7. A method according to claim 1 wherein said soybean plant that is selected in step (0) contains an endogenous

palmitic acid content in the oil of the mature seed formed

thereon of at least about 30% by weight as determined by

gas chromatography.

8. A method according to claim 1 wherein said soybean plant that is selected in step (c) contains an endogenous

palmitic acid content in the oil of the mature seed formed thereon from about 21% to about 30% by weight as deter

mined by gas chromatography.

9. A mature soybean seed formed by the self pollination of the plant selected in step (c) of the method of claim 1 and descendants thereof which upon self pollination.

10. A mature soybean seed according to claim 3 that contains an endogenous palmitic acid content in the oil thereof of at least about 18% by weight as determined by gas

chromatography.

11. A mature soybean seed according to claim 3 that contains an endogenous palmitic acid content in the oil thereof of greater than about 20% by weight as determined

by gas chromatography.

12. A mature soybean seed according to claim 3 that

contains an endogenous palmitic acid content in the oil thereof of greater than about 25% by weight as determined

by gas chromatography.

13. A mature soybean seed according to claim 3 that contains an endogenous palmitic acid content in the oil thereof of at least about 30% by weight as determined by gas

chromatography.

14. A mature soybean seed according to claim 3 that contains an endogenous palmitic acid content in the oil thereof from about 21% to about 30% by weight as deter

mined by gas chromatography.

(9)

UNITED STATES PATENT AND

TRADEMARK

OFFICE

CERTIFICATE OF CORRECTION

PATENT NO. ;

5,602,311

DATED

1

February 11, 1997

'NVENTOR<S> ‘

WALTER R. FEHR et a1.

It is certified that error appears in the above-indenti?ed patent and that said Letters Patent is hereby

corrected as shown below:

On the title page and column 1, line 1

IN THE TITLE [54]:

Please delete "SOYBEANS", second occurrence, and insert --SOYBEAN—-.

Signed and Sealed this

Third Day of June, 1997

BRUCE LEHMAN

Figure

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